Individual tumor volumes as function of time.

<p>Individual raw data measurements of tumor volumes for SKOV-3 (A) and SKBR-3 (B) xenografts in mice. Panel labels: Groups labeled 1000 and 4×250 are given ²²⁷Th-trastuzumab at dosages of 1000 kBq/kg as single injection or 4 · 250 kBq/kg at intervals marked (d  =  days or w  =  weeks).</p

Therapeutic effect of single injection and multiple injections of ²²⁷Th-trastuzumab.

*<p>Number of tumors reaching 10-fold size (total number of tumors in therapy group).</p>†<p>Days (mean ± standard error; estimation is limited to the largest time to reach endpoint within group if it is censored).</p>‡<p>Number of days delayed in reaching 10-fold increase in size compared to saline control group (mean ± standard error).</p>§<p>Significantly different from saline control group (p<0.001 for SKOV-3 and SKBR-3).</p>∥<p>Significantly different from trastuzumab control group (p<0.01 for SKOV-3 and p<0.001 for SKBR-3).</p

Normalized average tumor volumes as function of time.

<p>Averages of normalized tumor volumes for pilot experiment in SKBR-3 tumor xenografts (A) and further experiments in SKOV-3 (B) and SKBR-3 (C) tumor-carrying mice as function of days after start of therapy. Normalization was done by assigning all individual volumes at day 0 to the average tumor volume within each experiment. Tumor volumes at day of termination for each mouse were kept in the analysis until all mice in the therapy group were killed. Error bars are standard error.</p

Dose rate and accumulated dose.

<p>Calculated dose rate based on measured radioactivity in selected organs in mice carrying SKOV-3 (A) and SKBR-3 (B) tumor xenografts. Dose rates are derived from biodistribution data presented previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042345#pone.0042345-Abbas1" target="_blank">[12]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042345#pone.0042345-Abbas2" target="_blank">[13]</a>. Cumulative dose calculations for SKOV-3 and SKBR-3 mice are shown for single injection with 1000 kBq/kg body weight and for multiple injections of 250 kBq/kg separated by 2 and 4 weeks in tumor (C, D). Calculations are based on the assumption that the shape of the dose rate curve is identical for every new injection.</p

Autoradiography images of SKOV-3 and SKBR-3 tumors.

<p>Autoradiography images of the radioactivity distribution in 5 µm thick frozen tissue sections of SKOV-3 (A–B) and SKBR-3 (C–D) tumor xenografts from mice given 600 kBq/kg body weight ²²⁷Th-trastuzumab 8 days prior to tumor excition. Films were exposed for 7 days in −80°C prior to development.</p

White blood cell counts as function of time.

<p>Normalized, pooled white blood cell counts for mice carrying SKOV-3 and SKBR-3 tumor xenografts as function of weeks after onset of therapy. In animals given saline (NaCl) data from animals without tumor are pooled with data from tumor-carrying animals. Whole blood from vena saphena lateralis was collected in EDTA-coated tubes and analyzed using an automated hematology analyzer (Scil Vet abc animal blood counter). Error bars are standard deviation. Results statistically significantly lower than baseline value are marked by an asterisk.</p

Survival with largest tumor diameter less than 16 mm.

<p>Survival of mice carrying SKOV-3 (A) and SKBR-3 (B) tumor xenografts as function of days after onset of therapy. The survival end point was tumor size of 16 mm diameter. Mice euthanized due to loss of weight were censored in each plot. Groups significantly different from saline (NaCl) control group is marked with an asterisk.</p

CD146 expression in human osteosarcoma cell lines.

<p>A representative flow cytometry histogram that compare the binding of murine anti-CD146 OI-3 antibody to three different human osteosarcoma cell lines: OHS (A), Saos-2 (B) and KPDX (C). Control samples are unstained cells and cells stained only with the FITC-conjugated secondary antibody (ab). In addition, the binding of murine anti-CD37 antibody HH1 was examined for OHS (A) and Saos-2 (B) cells. The samples were analyzed on a BD FacsCalibur.</p

Comparison of the binding ability of chimeric antibodies targeting different antigens on OHS cells.

<p>A representative flow cytometry histogram which shows binding of the chimeric versions of OI-3 to OHS cells, compared with samples incubated with cetuximab, trastuzumab and rituximab. Control samples are unstained cells and cells stained only with the FITC-conjugated secondary antibody (ab). The samples were run on a Guava EasyCyte HT. Take note that the samples with chimeric antibodies were run on a different platform, with different settings and secondary antibody, and the fluorescence intensity levels are therefore not directly comparable to runs with murine OI-3 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0165382#pone.0165382.g001" target="_blank">Fig 1</a>.</p

Biodistribution of ¹²⁵I-labeled antibodies in mice bearing OHS xenografts.

<p>Tumor to normal tissue ratios 24 hours after injection of ¹²⁵I-labeled antibodies in female nude mice bearing OHS tumor xenografts. Four to five animals were used for each radioimmunoconjugate, which gave from four to seven tumors per group. Error bars correspond to standard error of the mean.</p